Pyrophosphatase

Pyrophosphatase Inorganic, Thermostable forms orthophosphate by catalyzing the hydrolysis of inorganic pyrophosphate.  Lucigen’s Pyrophosphatase Inorganic, Thermostable is purified from an E. coli strain carrying a plasmid encoding pyrophosphatase from a Thermus species. It is supplied at a concentration of 2,000 U/ml.

References:
1. Deutscher, M.P. & Kornberg, A.J. (1969) J. Biol. Chem. 244, 3019-3028.
2. Tabor, S. & Richardson, C. (1990) J. Biol. Chem. 265, 8322-8328.

Storage:
The enzyme should be stored at -20°C and is stable for 12 months if handled properly.
Storage Buffer:
20 mM Tris-HCl (pH 8.0), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT and 50% glycerol.
Purity:
>99% pure by SDS-PAGE. No detectable DNA contamination.
Activity Determination:
One unit is the amount of enzyme that will generate 1µmol of phosphate per minute from Pyrophosphatase Inorganic, Thermostable in a 10 minute reaction at 75°C in 50mM Tricine (pH 8.5), 1 mM MgCl₂ 0.32 mM PPi, in a reaction volume of 0.5 ml.
Absence of Endonuclease or Nicking Activity:
Incubation of 10 U of Pyrophosphatase Inorganic, Thermostable, with 1 µg of supercoiled pBR322 DNA for 16 hours at 70°C resulted in no detectable conversion to relaxed or linear forms by agarose gel electrophoresis.
Absence of Exonuclease Activity:
Incubation of 10 U of Pyrophosphatase Inorganic, Thermostable, with 1 µg of HindIII-cut lambda DNA  for 16 hours at 37°C and 65°C resulted in no smearing of bands on agarose gels.
Heat Inactivation: No